Dataset Information

Reliable blood cancer cells' telomere length evaluation by qPCR.

ABSTRACT:

Background

Telomere shortening is linked to a range of different human diseases, hence reliable measurement methods are needed to uncover such associations. Among the plethora of telomere length measurement methods, qPCR is reported as easy to conduct and a cost-effective approach to study samples with low DNA amounts.

Methods

Cancer cells' telomere length was evaluated by relative and absolute qPCR methods.

Results

Robust and reproducible telomere length measurements were optimized taking into account a careful reference gene selection and by knowing the cancer cells ploidy. qPCR data were compared to "gold standard" measurement from terminal restriction fragment (TRF).

Conclusions

Our study provides guidance and recommendations for accurate telomere length measurement by qPCR in cancer cells, taking advantage of our expertise in telomere homeostasis investigation in primary cutaneous T-cell lymphomas. Furthermore, our data emphasize the requirement of samples with both, high DNA quality and high tumor cells representation.

SUBMITTER: Ropio J

PROVIDER: S-EPMC7196062 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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Publications

Reliable blood cancer cells' telomere length evaluation by qPCR.

Ropio Joana J Chebly Alain A Ferrer Jacky J Prochazkova-Carlotti Martina M Idrissi Yamina Y Azzi-Martin Lamia L Cappellen David D Pham-Ledard Anne A Soares Paula P Merlio Jean-Philippe JP Chevret Edith E

Cancer medicine 20200306 9

<h4>Background</h4>Telomere shortening is linked to a range of different human diseases, hence reliable measurement methods are needed to uncover such associations. Among the plethora of telomere length measurement methods, qPCR is reported as easy to conduct and a cost-effective approach to study samples with low DNA amounts.<h4>Methods</h4>Cancer cells' telomere length was evaluated by relative and absolute qPCR methods.<h4>Results</h4>Robust and reproducible telomere length measurements were ...[more]

PMID: 32142223

Similar Datasets

Project description:BackgroundTelomeres, the protective cap of chromosomes, have emerged as powerful markers of biological age and life history in model and non-model species. The qPCR method for telomere length estimation is one of the most common methods for telomere length estimation, but has received recent critique for being too error-prone and yielding unreliable results. This critique coincides with an increasing awareness of the potentials and limitations of the qPCR technique in general and the proposal of a general set of guidelines (MIQE) for standardization of experimental, analytical, and reporting steps of qPCR. In order to evaluate the utility of the qPCR method for telomere length estimation in non-model species, we carried out four different qPCR assays directed at humpback whale telomeres, and subsequently performed a rigorous quality control to evaluate the performance of each assay.ResultsPerformance differed substantially among assays and only one assay was found useful for telomere length estimation in humpback whales. The most notable factors causing these inter-assay differences were primer design and choice of using singleplex or multiplex assays. Inferred amplification efficiencies differed by up to 40% depending on assay and quantification method, however this variation only affected telomere length estimates in the worst performing assays.ConclusionOur results suggest that seemingly well performing qPCR assays may contain biases that will only be detected by extensive quality control. Moreover, we show that the qPCR method for telomere length estimation can be highly precise and accurate, and thus suitable for telomere measurement in non-model species, if effort is devoted to optimization at all experimental and analytical steps. We conclude by highlighting a set of quality controls which may serve for further standardization of the qPCR method for telomere length estimation, and discuss some of the factors that may cause variation in qPCR experiments.

Dataset Information

Reliable blood cancer cells' telomere length evaluation by qPCR.

Background

Methods

Results

Conclusions

Publications

Reliable blood cancer cells' telomere length evaluation by qPCR.

Similar Datasets

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